Introduction
Macrophages in the tumour microenvironment
M1-Classically Activated | M2-Alternatively Activated | TAMS | References | |
---|---|---|---|---|
Activation
| INFγ, LPS | IL-4, IL-13, IL-10 | CSF-1, VEGF, CCL2, CCL3, CCL4, CCL5, CCL8, MCP-1, IL-4, IL-13, IL-10, TGFβ-1, PGE2. | Coffelt et al [4] Joyce and Pollard [9] Mantovani et al [84] |
Membrane Receptors
| TLR2, TLR4, CD16,CD32, CD64, CD80, CD86 | Scavenger receptor A, Scavenger receptor B, CD14, CD23, CD163 | CD11b+, CD14-, CD31-, CD45+, CD68+, CD117-, CD122-, CD146-, CD204+, CD206+, CCR2+,CSF1R+, MHCII+, CD23+, CD163+, CXCR4+, VEGFR1+, VEGFR2-, F4/80+(mice) | |
Cytokines produced
| IL-1, IL-6, IL-12, TNF, RNI, ROI | IL-1ra, IL-1 decoy receptor, EGF, FGF, VEGF, TNF-β, | bFGF, FGF, HFG, EGFR, PDGF, VEGF, ANG1, ANG2, IL-1, IL-8, TNF-α, TP, MMP-2, MMP-2, MMP-9, NO, CSF-1 | |
Chemokines produced
| CCL-2, CCL-3, CCL-4, CCL-5 CXCL8, CXCL9, CXCL10, CXCL11 | CCL-12, CCL-16, CCL-17, CCL-18, CCL-22, CCL-24 | CCL-2, CCL-3 | |
Marker
| iNOS | Arginase | F4/80 (mice), CD34 (humans) |
Tumour associated Macrophages (TAMs)
The role of TAMs in angiogenesis
The role of TAMs in tumour growth
The roles of TAMs in migration, invasion and intravasation
The role of TAMs in metastasis
The role of TAMs in immunosuppression
Correlation of TAMs to prognosis and survival
Bisphosphonates
Classification
Mechanism of action of Nitrogen-containing Bisphosphonates
Pharmacokinetics
Bisphosphonate | Potency | Clinical Dose | Route | Ref. |
---|---|---|---|---|
Etidronate | 1 | Oral, IV | [59] | |
Clodronate | 10 | 1600 mg/day | Oral | |
Pamidronate | 100 | 90 mg. 3-4 weeks | IV | |
Alendronate | 1000 | 10 mg/kg oral. 1 mg/kg I.V) | Oral | |
Ibandronate | 1,000 - 10,000 | 6 mg 3-4 weeks | IV | [59] |
Risedronate | 1,000-10,000 | 30 mg/d | Oral | [59] |
Zoledronic acid | 100,000 | 4 mg/2-3 weeks | IV |
Effects of Bisphosphonates on Osteoclasts
Anti-tumour activity of N-BPs - examples from breast cancer
Effect on tumour cells in bone
Direct anti-tumour effects
Effects on tumour cell adhesion and invasion
Effects on apoptosis and proliferation
Effects of N-BPs on macrophages
Effects on apoptosis
Effects on proliferation
Inhibition of pro-angiogenic factors: MMP-9
Inhibition of pro-angiogenic factors in clinical studies
Reversing M2 polarisation
Effect of bisphosphonate-induced macrophage depletion on tumour growth in vivo
Cell Type | Bisphosphonate | Main Findings | Reference |
---|---|---|---|
J774 cells RAW 264 cells | PAM, ALN, IBA, 100 μM 24 hours | Inhibited macrophage proliferation. Reduced cell viability. Increased cell death. | Rogers et al [68] |
J774 cells | ALN, 25 μM or 100 μM IBA 5 μM, 7.5 μM or 10 μM 24 hours | Dose dependent increase in accumulation of unprenylated Rap1A. | Firth et al [97] |
RAW 264 cells | ALN 10 μM; 5, 7, 9 or 16 hours ALN 100 μM; 1, 3, 5, 7, 9, 16 hours. | Dose and time dependent increase in accumulation of unprenylated Rap1A. Detectable after 16 hours incubation with 10 μM or 5 hours incubation with 100 μM | Monkkonen et al [98] |
J774 A.1 cells | PAM, ZOL, ALN, RIS 1-100 μM for 72 hours. | All BPs induced significant apoptosis ZOL > RIS > ALN > PAM. | Moreau et al [49] |
Macrophage precursor from bone marrow cells and bone marrow derived macrophages | PAM 2.5 × 10-7M (=0.25 μM), 96 hours | Significant inhibition M-CSF induced proliferation of bone marrow precursors | Cecchini et al [69] |
Activated human monocyte/macrophage | PAM 100-300 μM 24 hour pre-treatment Then activated with LPS. | Dose-dependent inhibition of MMP-9 expression Lower doses PAM increases expression. | Valleala et al [71] |
Human macrophage-like cell line U937 | Clodrolip 20-200 μM 2 days | Decreased cell survival | Hiraoka et al [85] |
Murine Macrophages | Clodrolip 9 μg per well 29 hours | Decreased cell viability | Gazzangia et al [86] |
Murine peritoneal macrophages | Clodrolip 1 mg/ml 6 hours. | Dose-dependent increase in apoptosis. | Zeisberger et al [87] |
Bone marrow cells from naive mice cultured with M-CSF or tumour supernatant. | Zoledronic acid 0.03, 0.15, 0.3 μM 6 days | Dose-dependent inhibition in differentiation of myeloid cells to macrophages. Decreased in M2 phenotype compared to control. | Veltman et al [89] |
Model | Bisphosphonate | Main Findings | Reference |
---|---|---|---|
BALB-neut mice with mammary tumour virus (rat c-erb-2-neu/transgene) | ZOL 0.1 mg/kg or PAM 2 mg/kg 5 days a week. | Zol decreased macrophage infiltration into tumour stroma associated with decreased levels of pro-MMP-9 and VEGF | Melani C et al [21] |
Mammary carcinoma cells implanted in BALB-neutT mice | ZOL 100 μM/kg Once a week for 4 weeks, followed by 3 weeks rest, cycle continued. | Impaired TAM recruitment and infiltration into tumour and reduced neo-vascularisation reversal of TAM polarity from pro-tumoural M2 to tumoricidal M1 | Coscia et al [83] |
HARA-B lung cancer cells implanted in BALB/c nude mice | Clodrolip 200 μL or 400 μL Every 3 days for 6 weeks (s.c.). | Reduced TAM infiltration correlated to reduced metastatic spread | Hiraoka et al [85] |
Human melanoma cell line IIB-MEL-J with or without MCP-1 expression vector. Athymic male NIC-(S)-Nu mice | Clodrolip 50 μl or 200 μl (6 mg clodronate per 1 ml ) From day before cell injection and every 5 to 7 days thereafter. | Reduced TAMs infiltration correlated to decreased tumour volume and angiogenesis and increased survival | Gazzangia et al [86] |
F9 teratocarcinoma cells implanted in SV129 female mice | Clodrolip, 1 mg/20 g every 4 days. | Reduced TAM infiltration. 82% reduction in tumour volume | Zeisberger et al [87] |
A673 rhabdomyosarcoma cells into CD-1 nude mice | Clodrolip, 1 mg/20 g every 4 days. | 93% reduction in TAM 89% reduction in blood vessel density | Zeisberger et al [87] |
Metastatic liver cancer Mouse model LM3R or SMMC7721 human hepatocellular cell lines in BALB/c nu/nu mice | Clodrolip 100 μg/kg 3 times a week or, ZOL 100 μg/kg 3 times a week and or 30 mg/kg sorafenib. | Reduced TAM infiltration with combination therapy. Correlated with decreased tumour growth, angiogenesis and lung metastasis. ZOL had greater effect than clodrolip | Zhang et al [88] |
Cervical carcinoma K14-HPV16 transgenic mice | ZOL 100 μg Every day for 3 months. | Decreased MMP-9 expression by TAMs | Giraudo et al [72] |
Peritoneal macrophage obtained from CBA-J mice injected with AC29 mesothelioma cells | Clodronate 200 μl twice over 10 days. | Depleted peritoneal macrophages. Decreased tumour growth | Veltman et al [89] |
CBA-J mice injected with AC29 mesothelioma cells | ZOL 100 μg/kg s.c. Every day for 25 days. | Increased myeloid precursors. Decreased TAMs No significant increase in survival or decrease in tumour burden | Veltman et al [89] |